Studies of P-glycoprotein and drug interactions

P-糖蛋白和药物相互作用的研究

基本信息

批准号：
9230408
负责人：
Qinghai Zhang
金额：
$ 50.23万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9230408
关键词：
ATP phosphohydrolase ATP-Binding Cassette Transporters Address Affect Affinity Antibody-drug conjugates Antineoplastic Agents Binding Biological Assay Biological Availability Blood - brain barrier anatomy Bos taurus structural-GP protein Brain Cells Central Nervous System Diseases Cessation of life Chemicals Chemistry Clinical Collaborations Complex Crystallization Detergents Development Disease Dolastatin Compound Drug Design Drug Efflux Drug Industry Drug Interactions Drug Kinetics Drug or chemical Tissue Distribution Drug resistance Drug usage Electron Microscopy Energy Transfer Epilepsy Evaluation Excretory function Exhibits FDA approved Genetic Goals Grant HIV Human Intestines Investigational Drugs Kidney Kinetics Libraries Ligand Binding Ligands Lipids Literature Liver Location Malignant Neoplasms Mediating Membrane Transport Proteins Molecular Conformation Multi-Drug Resistance P-Glycoprotein P-glycoprotein 2 Patients Pattern Peptides Pharmaceutical Preparations Pharmacologic Substance Pharmacology Phenotype Positioning Attribute Predisposition Research Personnel Resistance Resolution Roentgen Rays Route Side Structural Biologist Structure Structure-Activity Relationship Techniques Testing Time Treatment Efficacy Vinca Alkaloids X-Ray Crystallography absorption analog base biophysical techniques chemical property chemical synthesis chemotherapy clinical efficacy clinically significant design drug candidate drug discovery drug efficacy drug market flexibility glycoprotein structure human disease indium arsenide inhibitor/antagonist lipophilicity luminance novel novel therapeutics particle preclinical study public health relevance scaffold small molecule ward

项目摘要

DESCRIPTION (provided by applicant): P-glycoprotein (Pgp) is a highly dynamic ATP-binding cassette (ABC) membrane transporter that effluxes a diversity of molecules out of cells. It is a major determinant of drug absorption, distribution, and excretion in intestines, liver, kidney and brain, and also an impediment to successful chemotherapy in some cancers, HIV and central nervous system (CNS) diseases. About half of marketed drugs are estimated to be transport substrates or inhibitors of Pgp, and the evaluation of Pgp susceptibility of drug candidates has become an important step in the development of new therapeutics in the pharmaceutical industry. However, Pgp binding molecules exhibit complex activities, acting as ATPase stimulators, inhibitors, transport substrates, or non- transportable ligands. Despite extensive studies, we still have very limited understanding of the mechanisms for the complex and polyspecific Pgp-drug interactions pertaining to Pgp transport, inhibition and evasion. This study aims to (1) define how structural and chemical properties of a ligand affect its interactions with Pgp, (2) characterize how Pgp reacts upon the binding of different classes of ligands, and (3) rationalize chemical synthesis to modify existing drugs to evade Pgp transport. We will use an integrative chemistry, structural and functional approach to tackle these aims, which is based on several major advances in the literature and that we have most recently made on Pgp structural determination and ligand interactions. First, we propose to conduct a thorough structure-activity relationship study within a focused library of ligands bearing common scaffolds. We will use a battery of functional assays, including ATPase activity, detailed drug binding and competition, cell-based transport, and drug resistance assays, as well as structural characterizations to evaluate these compounds. Second, we will use several complementary biophysical techniques, including X- ray crystallography, single particle electron microscopy (EM), and luminance resonance energy transfer (LRET), for the determination of Pgp/ligand complex structures, conformational distributions, and the kinetics of conformational changes. The characterization of Pgp conformations by EM and LRET, together with the screen of novel ligands, detergents and lipids, as well as new constructs to stabilize Pgp (and certain conformations), will facilitate higher resolution (< 3.0 Å) structural studies, a critical barrier that has eluded Pgp thus far. Third, we will modify several Pgp drug substrates on positions that have been identified without diminution of drug potency, thus with a focus on resulting changes in Pgp interactions. By the related three aim studies we will achieve a detailed and fundamental understanding of polyspecific Pgp-drug interactions, which will have a far- reaching impact on drug discovery given the pharmacological and clinical significance of Pgp and that many current and investigational drugs are susceptible to Pgp efflux.

描述（由申请人提供）：P-糖蛋白（Pgp）是一种高度动态的 ATP 结合盒（ABC）膜转运蛋白，可将多种分子排出细胞外，它是药物吸收、分布和排泄的主要决定因素。据估计，大约一半的上市药物会被运输到肠道、肝脏、肾脏和大脑，并且还会阻碍某些癌症、艾滋病毒和中枢神经系统 (CNS) 疾病的成功化疗。然而，Pgp 结合分子表现出复杂的活性，可作为 ATP 酶刺激剂、抑制剂、转运底物、尽管进行了广泛的研究，但我们对与 Pgp 转运、抑制和逃避有关的复杂和多特异性 Pgp-药物相互作用的机制的了解仍然非常有限。旨在 (1) 定义配体的结构和化学性质如何影响其与 Pgp 的相互作用，(2) 表征 Pgp 在不同类别配体的结合上如何反应，以及 (3) 合理化化学合成以修改现有药物以逃避 Pgp我们将使用综合化学、结构和功能方法来实现这些目标，该方法基于文献中的几项重大进展以及我们最近在 Pgp 结构测定和配体相互作用方面取得的进展。彻底我们将使用一系列功能测定，包括 ATP 酶活性、详细的药物结合和竞争、基于细胞的转运和耐药性测定，以及结构表征来进行结构-活性关系研究。其次，我们将使用几种互补的生物物理技术，包括 X 射线晶体学、单粒子电子显微镜 (EM) 和亮度共振能量转移 (LRET)，来确定 Pgp/配体复合物结构，通过 EM 和 LRET 表征 Pgp 构象，以及新配体、去污剂和脂质以及稳定 Pgp（和某些构象）的新结构的筛选，将有助于提高分辨率。 (< 3.0 Å) 结构研究，这是迄今为止 Pgp 未能解决的一个关键障碍。第三，我们将在未经鉴定的位置上修改几种 Pgp 药物底物。药物效力的降低，因此重点关注 Pgp 相互作用所产生的变化，通过相关的三个目标研究，我们将对多特异性 Pgp-药物相互作用有一个详细和基本的了解，这将对药物发现产生深远的影响。 Pgp 的药理学和临床意义以及许多当前和研究中的药物对 Pgp 外流敏感。